Assay Technologies for Proteases

Roy, Anuradha; Lushington, Gerald H.; McGee, J; Chaguturu, Rathnam

doi:10.1002/9781118739907.ch1

Cited by 3 publications

(2 citation statements)

References 138 publications

(264 reference statements)

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“…Fluorimetric MMP peptides have been previously used to measure and quantify MMP activities from dentin and oral bacteria as they have higher sensitivity than colorimetric assays [15,17,35]. In the current study, human neutrophils showed activity towards the MMP substrates, and the activities either remained constant or became elevated throughout the 48 hours replacement cycle, suggesting their potential ability to degrade the collagen type I in tooth dentin.…”

Section: Mmp Activities From Neutrophilsmentioning

confidence: 57%

Human neutrophils degrade methacrylate resin composites and tooth dentin

et al. 2019

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Cholesterol esterase-like (CE) activity from saliva and esterase from cariogenic bacteria hydrolyze ester linkages of dental methacrylate resins. Collagenolytic, matrix metalloproteinase-like (MMP) activities from dentin and bacteria degrade collagen in demineralized tooth dentin. Human neutrophils in the oral cavity contain factors that are hypothesized to have CE and MMP activities that could contribute to the degradation of methacrylate resins and dentinal collagen. Objectives: To measure the CE and MMP activities from human neutrophils and their ability to degrade dental methacrylate resin composite and dentinal collagen.

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Section: Mmp Activities From Neutrophilsmentioning

confidence: 57%

Human neutrophils degrade methacrylate resin composites and tooth dentin

et al. 2019

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“…From these energy predictions, it is reasonable to infer that the exothermic energy gained from Spike binding to TMPRSS2, MSP, and PSP may, in all three cases, provide ample energetic impetus for the enzyme to overcome the reactive barrier and cleave the peptide. While the precise reaction kinetics have not been reported for proteolytic cleavage by MSP and PSP, it is worth noting that widespread conservation of protease catalytic triad structures imposes homogeneity on reactive proficiency across the broad manifold of serine proteases [229][230][231]. Known proteolysis barriers for HIV protease (15.1-17.9 kcal/mol) [232,233], for example, bear a strong resemblance to TMPRSS2 reactive barriers.…”

Section: Prospective Biochemical Rationalizationsmentioning

confidence: 99%

Bacterial Proteases as Potentially Exploitable Modulators of SARS-CoV-2 Infection: Logic from the Literature, Informatics, and Inspiration from the Dog

Lushington,

Linde,

Melgarejo

2023

BioTech

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(1) Background: The COVID-19 pandemic left many intriguing mysteries. Retrospective vulnerability trends tie as strongly to odd demographics as to exposure profiles, genetics, health, or prior medical history. This article documents the importance of nasal microbiome profiles in distinguishing infection rate trends among differentially affected subgroups. (2) Hypothesis: From a detailed literature survey, microbiome profiling experiments, bioinformatics, and molecular simulations, we propose that specific commensal bacterial species in the Pseudomonadales genus confer protection against SARS-CoV-2 infections by expressing proteases that may interfere with the proteolytic priming of the Spike protein. (3) Evidence: Various reports have found elevated Moraxella fractions in the nasal microbiomes of subpopulations with higher resistance to COVID-19 (e.g., adolescents, COVID-19-resistant children, people with strong dietary diversity, and omnivorous canines) and less abundant ones in vulnerable subsets (the elderly, people with narrower diets, carnivorous cats and foxes), along with bioinformatic evidence that Moraxella bacteria express proteases with notable homology to human TMPRSS2. Simulations suggest that these proteases may proteolyze the SARS-CoV-2 spike protein in a manner that interferes with TMPRSS2 priming.

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Endodontic pathogens possess collagenolytic properties that degrade human dentine collagen matrix

et al. 2018

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Aim To measure collagenolytic protease activity from Enterococcus faecalis and Micrococcus luteus and their ability to degrade human dentinal collagen. Methodology Proteases activity of E. faecalis ATCC 29212, ATCC 47077 and M. luteus towards generic and specific human matrix metalloproteinase (MMP) substrates was measured using a fluorimetric assay. The ability of the bacteria to degrade dentinal collagen was tested by quantifying the amount of hydroxyproline released into the media following incubation of the bacteria or heat‐inactivated bacteria (HIN) with demineralized human dentine samples for 24 h and by scanning electron microscopy (SEM). Multifactorial anova and Tukey's post hoc test were used to analyse the data (P < 0.05). Results All strains had MMP‐like activities, but with different substrate affinity; E. faecalis ATCC 29212, ATCC 47077 and M. luteus had the greatest affinity towards MMP‐8 (7.75 ± 0.88 μmol L−1/3 × 106 CFU), MMP‐9 (33.86 ± 5.16 μmol L−1/3 × 106 CFU) and generic MMP (26.08 ± 4.48 μmol L−1/3 × 106 CFU), respectively. The amount of hydroxyproline released from demineralized dentine was similar (P > 0.05) for the three strains (range 1.8 ± 0.17 to 2.38 ± 0.39 μg 50 μL−1) and was significantly higher (P < 0.001) compared to their HIN counterparts (0.61 ± 0.22 μg 50 μL−1). SEM revealed increased collagen network degradation after incubation with bacteria versus HIN. Conclusions Endodontic pathogens possess collagenolytic protease properties that enable them to degrade dentinal collagen, potentially compromising the restoration‐tooth and sealer‐tooth interfaces. These collagenolytic protease properties could facilitate the migration of pathogenic bacteria into the root canal system and explain in part their role in root canal infections.

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Assay Technologies for Proteases

Cited by 3 publications

References 138 publications

Human neutrophils degrade methacrylate resin composites and tooth dentin

Human neutrophils degrade methacrylate resin composites and tooth dentin

Bacterial Proteases as Potentially Exploitable Modulators of SARS-CoV-2 Infection: Logic from the Literature, Informatics, and Inspiration from the Dog

Endodontic pathogens possess collagenolytic properties that degrade human dentine collagen matrix

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